Complex interactions between human myoblasts and the surrounding 3D fibrin-based matrix.

Anchorage of muscle cells to the extracellular matrix is crucial for a range of fundamental biological processes including migration, survival and differentiation. Three-dimensional (3D) culture has been proposed to provide a more physiological in vitro model of muscle growth and differentiation tha...

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Autores principales:	Stéphane Chiron, Carole Tomczak, Alain Duperray, Jeanne Lainé, Gisèle Bonne, Alexandra Eder, Arne Hansen, Thomas Eschenhagen, Claude Verdier, Catherine Coirault
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/49058a5fb7ff4effadd06566ab2c3ffc
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spelling	oai:doaj.org-article:49058a5fb7ff4effadd06566ab2c3ffc2021-11-18T07:20:24ZComplex interactions between human myoblasts and the surrounding 3D fibrin-based matrix.1932-620310.1371/journal.pone.0036173https://doaj.org/article/49058a5fb7ff4effadd06566ab2c3ffc2012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22558372/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Anchorage of muscle cells to the extracellular matrix is crucial for a range of fundamental biological processes including migration, survival and differentiation. Three-dimensional (3D) culture has been proposed to provide a more physiological in vitro model of muscle growth and differentiation than routine 2D cultures. However, muscle cell adhesion and cell-matrix interplay of engineered muscle tissue remain to be determined. We have characterized cell-matrix interactions in 3D muscle culture and analyzed their consequences on cell differentiation. Human myoblasts were embedded in a fibrin matrix cast between two posts, cultured until confluence, and then induced to differentiate. Myoblasts in 3D aligned along the longitudinal axis of the gel. They displayed actin stress fibers evenly distributed around the nucleus and a cortical mesh of thin actin filaments. Adhesion sites in 3D were smaller in size than in rigid 2D culture but expression of adhesion site proteins, including α5 integrin and vinculin, was higher in 3D compared with 2D (p<0.05). Myoblasts and myotubes in 3D exhibited thicker and ellipsoid nuclei instead of the thin disk-like shape of the nuclei in 2D (p<0.001). Differentiation kinetics were faster in 3D as demonstrated by higher mRNA concentrations of α-actinin and myosin. More important, the elastic modulus of engineered muscle tissues increased significantly from 3.5 ± 0.8 to 7.4 ± 4.7 kPa during proliferation (p<0.05) and reached 12.2 ± 6.0 kPa during differentiation (p<0.05), thus attesting the increase of matrix stiffness during proliferation and differentiation of the myocytes. In conclusion, we reported modulations of the adhesion complexes, the actin cytoskeleton and nuclear shape in 3D compared with routine 2D muscle culture. These findings point to complex interactions between muscle cells and the surrounding matrix with dynamic regulation of the cell-matrix stiffness.Stéphane ChironCarole TomczakAlain DuperrayJeanne LainéGisèle BonneAlexandra EderArne HansenThomas EschenhagenClaude VerdierCatherine CoiraultPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 4, p e36173 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Stéphane Chiron Carole Tomczak Alain Duperray Jeanne Lainé Gisèle Bonne Alexandra Eder Arne Hansen Thomas Eschenhagen Claude Verdier Catherine Coirault Complex interactions between human myoblasts and the surrounding 3D fibrin-based matrix.
description	Anchorage of muscle cells to the extracellular matrix is crucial for a range of fundamental biological processes including migration, survival and differentiation. Three-dimensional (3D) culture has been proposed to provide a more physiological in vitro model of muscle growth and differentiation than routine 2D cultures. However, muscle cell adhesion and cell-matrix interplay of engineered muscle tissue remain to be determined. We have characterized cell-matrix interactions in 3D muscle culture and analyzed their consequences on cell differentiation. Human myoblasts were embedded in a fibrin matrix cast between two posts, cultured until confluence, and then induced to differentiate. Myoblasts in 3D aligned along the longitudinal axis of the gel. They displayed actin stress fibers evenly distributed around the nucleus and a cortical mesh of thin actin filaments. Adhesion sites in 3D were smaller in size than in rigid 2D culture but expression of adhesion site proteins, including α5 integrin and vinculin, was higher in 3D compared with 2D (p<0.05). Myoblasts and myotubes in 3D exhibited thicker and ellipsoid nuclei instead of the thin disk-like shape of the nuclei in 2D (p<0.001). Differentiation kinetics were faster in 3D as demonstrated by higher mRNA concentrations of α-actinin and myosin. More important, the elastic modulus of engineered muscle tissues increased significantly from 3.5 ± 0.8 to 7.4 ± 4.7 kPa during proliferation (p<0.05) and reached 12.2 ± 6.0 kPa during differentiation (p<0.05), thus attesting the increase of matrix stiffness during proliferation and differentiation of the myocytes. In conclusion, we reported modulations of the adhesion complexes, the actin cytoskeleton and nuclear shape in 3D compared with routine 2D muscle culture. These findings point to complex interactions between muscle cells and the surrounding matrix with dynamic regulation of the cell-matrix stiffness.
format	article
author	Stéphane Chiron Carole Tomczak Alain Duperray Jeanne Lainé Gisèle Bonne Alexandra Eder Arne Hansen Thomas Eschenhagen Claude Verdier Catherine Coirault
author_facet	Stéphane Chiron Carole Tomczak Alain Duperray Jeanne Lainé Gisèle Bonne Alexandra Eder Arne Hansen Thomas Eschenhagen Claude Verdier Catherine Coirault
author_sort	Stéphane Chiron
title	Complex interactions between human myoblasts and the surrounding 3D fibrin-based matrix.
title_short	Complex interactions between human myoblasts and the surrounding 3D fibrin-based matrix.
title_full	Complex interactions between human myoblasts and the surrounding 3D fibrin-based matrix.
title_fullStr	Complex interactions between human myoblasts and the surrounding 3D fibrin-based matrix.
title_full_unstemmed	Complex interactions between human myoblasts and the surrounding 3D fibrin-based matrix.
title_sort	complex interactions between human myoblasts and the surrounding 3d fibrin-based matrix.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/49058a5fb7ff4effadd06566ab2c3ffc
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Complex interactions between human myoblasts and the surrounding 3D fibrin-based matrix.

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